MRI-based detection of alkaline phosphatase gene reporter activity using a porphyrin solubility switch
| Autor: | Yelena Emer, Gil G. Westmeyer, Jutta Lintelmann, Alan Jasanoff |
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| Přispěvatelé: | Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology. Department of Nuclear Science and Engineering, Westmeyer, Gil G., Emer, Yelena, Jasanoff, Alan Pradip |
| Předmět: |
Male
Porphyrins Clinical Biochemistry Contrast Media Endogeny Biology medicine.disease_cause Biochemistry Article Adenoviridae Rats Sprague-Dawley Genes Reporter Gene expression Drug Discovery medicine Animals Humans Protein Isoforms Gene Molecular Biology chemistry.chemical_classification Pharmacology HEK 293 cells Brain General Medicine Alkaline Phosphatase Magnetic Resonance Imaging Rats Enzyme HEK293 Cells chemistry Solubility Alkaline phosphatase Molecular Medicine Intracellular |
| Zdroj: | Europe PubMed Central PMC Chem. Biol. 21, 422-429 (2014) |
| Popis: | The ability to map patterns of gene expression noninvasively in living animals could have impact in many areas of biology. Reporter systems compatible with MRI could be particularly valuable, but existing strategies tend to lack sensitivity or specificity. Here we address the challenge of MRI-based gene mapping using the reporter enzyme secreted alkaline phosphatase (SEAP), in conjunction with a water-soluble metalloporphyrin contrast agent. SEAP cleaves the porphyrin into an insoluble product that accumulates at sites of enzyme expression and can be visualized by MRI and optical absorbance. The contrast mechanism functions in vitro, in brain slices, and in animals. The system also provides the possibility of readout both in the living animal and by postmortem histology, and it notably does not require intracellular delivery of the contrast agent. The solubility switch mechanism used to detect SEAP could be adapted for imaging of additional reporter enzymes or endogenous targets. Raymond and Beverley Sackler Foundation National Institutes of Health (U.S.) (New Innovator Award Grant DP2-OD2441) MIT-Germany Seed Fund (Grant) |
| Databáze: | OpenAIRE |
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